Antimicrobial effects of photodynamic therapy with Fotoenticine on Streptococcus mutans isolated from dental caries.

Photodynamic therapy (PDT) is a promising strategy to control cariogenic pathogens, such as Streptococcus mutans. Seeking to reach the total bacterial elimination from dental surfaces, novel photosensitizers have been investigated, such as Fotoenticine (FTC) derived from chlorin e6. The objective of this study was to investigate the photodynamic effects of FTC against several clinical strains of S. mutans. Clinical isolates were obtained from patients with active carious lesions, identified by molecular analysis and subjected to PDT using laser irradiation (660 nm and 39.5 J/cm2) in planktonic and biofilm stages. We identified 11 S. mutans strains from cervical, occlusal and proximal caries. PDT mediated by FTC has totally eliminated the S. mutans cells in planktonic growth for all analyzed strains. In biofilms, PDT with FTC reached statistically significant reductions compared with the non-treated control group, at 5.4, 5.5 and 6.5 Log10 (CFU/mL), respectively, for the strains from proximal, occlusal and cervical caries. The scanning electron microscopy evaluations confirmed that PDT mediated by FTC was able to disaggregate and kill the S. mutans cells adhered to enamel surface, suggesting its potential to disinfect the dental tissues.

Photodynamic Activity on Biofilm in Endotracheal Tubes of Patients Admitted to an Intensive Care Unit.

Ventilator-associated pneumonia (VAP) is an infection that arises after endotracheal intubation affecting patients under intensive care. The presence of the endotracheal tube (ETT) is a risk factor since it is colonized by multispecies biofilm. Antimicrobial photodynamic therapy (aPDT) could be a strategy to decontaminate ETTs. We verify if methylene blue (MB) associated with external illumination of the ETT could be an alternative to destroy biofilm. We performed an in vitro and ex vivo study. In vitro study was performed with P. aeruginosa biofilm grew over ETT for 7 days. After treatment, the surviving cells were cultured for 3 days and the biofilm was analyzed by crystal violet absorbance. Ex vivo study employed ETT obtained from extubated patients. aPDT was performed with MB (100 µm) and red LED (λ = 640±20 nm). We quantified the biofilm thickness and used scanning electron microscopy and fluorescence technique to verify morphological and functional changes after aPDT. Our results showed that bacteria remain susceptible to aPDT after sequential treatments. We also attested that aPDT can reduce biofilm thickness, disrupt biofilm attachment from ETT surface and kill microbial cells. These data suggest that aPDT should be investigated to decrease VAP incidence via ETT decontamination.

Comparative analysis of the effects of ozone and ultrasound on Streptococcus mutans

Streptococcus mutans is one of the principal pathogens of the human oral habitat, being one of the principal etiological agents of carious lesions. Ozone is a powerful oxidant, it has the ability to inactivate microorganisms in general, and ultrasound is an acoustic system generated through a piezoelectric crystal that also presents microbicidal effects. In the present study, a comparative analysis was made of the damage caused to Streptococcus mutans in vitro by ultrasound and ozone, through scanning electron microscopy (SEM) and atomic force microscopy (AF) analysis. In addition, flow cytometry was used to determine microbial viability and the formation of Reactive Oxygen Species (ROS) after the application of different techniques. The data obtained by means of micro- scopic analysis reveal that both ozone and ultrasound produce morphological alterations in bacteria, which become rod-shaped organisms. In addition to this deformation, on the microbial surface it was possible to identify crater-like impressions. In contrast, the irregularity of protuber- ances on the surface of the microbial wall was only detected when ozone was employed. Regarding the formation of ROS, it was observed that that ozone induces a significant growth (p < .05) of these molecules, while ultrasound does not present this effect. Ozone and ultrasound present micro- bicidal effects, however, ozone is more efficient.

Analysis of damage on the Streptococcus mutans immersed in ozonated water: preliminary study for application as mouth rinse

Ozonated water has been demonstrated to induce significant results in terms of the elimination ofmicroorganisms. The present study assessed the damage toStreptococcusmutansafter exposureto ozonated water; the ozone generator was adjusted to provide an outlet concentration of60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanningelectron microscopy and atomic force images were obtained to identify damage to the bacteria,followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed asignificant reduction in viability and the images evidenced the generation of gaps on themicrobial wall and surface layer alterations. Ozone can induce significant damage toS.mutans,thus suggesting that the use of ozonated water to prevent carious lesion formation is extremelypromising.

Analysis of damage on the Streptococcus mutans immersed in ozonated water: preliminary study for application as mouth rinse

Ozonated water has been demonstrated to induce significant results in terms of the elimination ofmicroorganisms. The present study assessed the damage toStreptococcusmutansafter exposureto ozonated water; the ozone generator was adjusted to provide an outlet concentration of60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanningelectron microscopy and atomic force images were obtained to identify damage to the bacteria,followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed asignificant reduction in viability and the images evidenced the generation of gaps on themicrobial wall and surface layer alterations. Ozone can induce significant damage toS.mutans,thus suggesting that the use of ozonated water to prevent carious lesion formation is extremelypromising.

Photodynamic damage predominates on different targets depending on cell growth phase of Candida albicans.

Photodynamic inactivation (PDI) has been reported to be effective to eradicate a wide variety of pathogens, including antimicrobial-resistant microorganisms. The aim of this study was to identify the potential molecular targets of PDI depending on growth phase of Candida albicans. Fungal cells in lag (6h) and stationary (48h) phases were submitted to PDI mediated by methylene blue (MB) combined with a (662±21) nm-LED, at 360mW of optical power. Pre-irradiation time was 10min and exposure times were 12min, 15min and 18min delivering radiant exposures of 129.6J/cm2, 162J/cm2 and 194.4J/cm2, respectively, on a 24-well plate of about 2cm2 at an irradiance of 180mW/cm2. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force spectroscopy (AFS) and Fourier transform infrared spectroscopy (FT-IR) were employed to evaluate the photodynamic effect in young and old fungal cells following 15min of irradiation. Morphological analysis revealed wrinkled and shrunk fungal cell membrane for both growth phases while extracellular polymeric substance (EPS) removal was only observed for old fungal cells. Damaged intracellular structures were more pronounced in young fungal cells. The surface nanostiffness of young fungal cells decreased after PDI but increased for old fungal cells. Cellular adhesion force was reduced for both growth phases. Fungal cells in lag phase predominantly showed degradation of nucleic acids and proteins, while fungal cells in stationary phase showed more pronounced degradation of polysaccharides and lipids. Taken together, our results indicate different molecular targets for fungal cells in lag and stationary growth phase following PDI.

Antimicrobial and anti-biofilm properties of polypropylene meshes coated with metal-containing DLC thin films.

A promising strategy to reduce nosocomial infections related to prosthetic meshes is the prevention of microbial colonization. To this aim, prosthetic meshes coated with antimicrobial thin films are proposed. Commercial polypropylene meshes were coated with metal-containing diamond-like carbon (Me-DLC) thin films by the magnetron sputtering technique. Several dissimilar metals (silver, cobalt, indium, tungsten, tin, aluminum, chromium, zinc, manganese, tantalum, and titanium) were tested and compositional analyses of each Me-DLC were performed by Rutherford backscattering spectrometry. Antimicrobial activities of the films against five microbial species (Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus faecalis) were also investigated by a modified Kirby-Bauer test. Results showed that films containing silver and cobalt have inhibited the growth of all microbial species. Tungsten-DLC, tin-DLC, aluminum-DLC, zinc-DLC, manganese-DLC, and tantalum-DLC inhibited the growth of some strains, while chromium- and titanium-DLC weakly inhibited the growth of only one tested strain. In-DLC film showed no antimicrobial activity. The effects of tungsten-DLC and cobalt-DLC on Pseudomonas aeruginosa biofilm formation were also assessed. Tungsten-DLC was able to significantly reduce biofilm formation. Overall, the experimental results in the present study have shown new approaches to coating polymeric biomaterials aiming antimicrobial effect.

AFM characterization of bovine enamel and dentine after acid-etching.

Teeth are constituted mainly of hydroxyapatite molecules (Ca(10)(PO(4))(6)(OH)(2)), grouped in different microstructural arrangements, depending on the dental layer considered (enamel or dentine). In the present work, these dental microstructural arrangements were characterized by atomic force microscopy. Enamel and dentine samples were cut from freshly extracted bovine incisor teeth. After metallographic polishing, the dental surfaces were etched with lactic acid (113.8 mmol/L, pH 3.3). Three etching times were tested: 1, 3 and 5 min. Atomic force micrographs showed that 1 min of etching time was effective to remove the smear layer, polishing debris and scratches, and display the characteristics of interest for both enamel and dentine. Although the bovine dental enamel rod cross-section presented keyhole-like shape, its measured dimensions (8.8 microm of major axis and 3.7 microm of minor axis) exhibited an insignificant discrepancy from human prisms diameters. Bovine dentinal tubules displayed larger mean diameters (4.0 microm) and a lower density (approximately 17,100 tubules/mm(2)) than human dentine, suggesting that the use of bovine dentine as a substitute for human dentine in resin adhesion investigations should be reconsidered. Apatite nanoparticles presented a mean radius (22-23 nm) considerably smaller than that of human teeth.

Color and surface temperature variation during bleaching in human devitalized primary teeth: an in vitro study.

This study's purpose was to make an in vitro assessment of 2 whitening techniques in primary teeth, regarding color and temperature surface variation, during dental bleaching using different catalytic sources. Twenty-one extracted human upper central deciduous incisors were used in this in vitro study. The teeth were darkened with human blood for a period of 21 days. After preparing the teeth, they were randomly distributed into 2 groups, according to bleaching source of activation: (1) a diode laser (DL) group; and (2) a halogen lamp (HL) group. The bleaching process was performed, according to the manufacturer's guidelines, using Whiteness HP (FGM, Joinville, Brazil). The color was assessed by spectrophotometer (CIELab) and the VITA scale (3M) before and immediately after tooth whitening. The temperature increase in the radicular surface during the bleaching was registered with a thermographic camera ThermaCAM SC 3000 (Flir Systems, Danderyd, Sweden) at the Nuclear and Energy Research Institute, IPEN-CNEN (São Paulo, Brazil). There was no significant difference between the groups in terms of color changes, but there was a statistically significant difference for temperature variation. The use of a diode laser and halogen lamp both promoted whitening in devitalized primary teeth in vitro. As a catalytic source of energy, the diode laser--with the applied parameters--promoted a smaller temperature increase compared to the halogen lamp during the bleaching procedure on nonvital primary teeth.